| 1996 |
PXAAA1 (PEX6) encodes a cytoplasmic AAA-family ATPase required for peroxisomal matrix protein import; substitution of arginine for the conserved lysine in the ATPase domain abolished biological activity, confirming ATPase activity is essential. PEX6 is also required for stability of the PTS1 receptor Pxr1p (PEX5). |
Functional complementation in patient fibroblasts, ATPase domain mutagenesis (K→R), immunolocalization, receptor stability assay |
The EMBO journal |
High |
8670792
|
| 1998 |
PEX1 and PEX6 physically interact with each other in vitro and in the yeast two-hybrid system; overexpression of PEX6 suppresses certain PEX1-deficient phenotypes in an allele-specific manner requiring partial PEX1 activity, and the common disease mutation PEX1-G843D attenuates the PEX1–PEX6 interaction. |
Yeast two-hybrid, in vitro binding (pull-down), allele-specific genetic suppression (overexpression rescue) |
Proceedings of the National Academy of Sciences of the United States of America |
High |
9671729
|
| 2011 |
AWP1/ZFAND6 interacts with PEX6 AAA ATPase (but not with PEX1–PEX6 complexes) and preferentially binds cysteine-monoubiquitinated PEX5 via its A20 zinc-finger domain; AWP1 stimulates PEX5 export in an in vitro assay and its knockdown impairs PTS1-protein import and reduces PEX5 stability, identifying AWP1 as a novel PEX6 cofactor in PEX5 recycling. |
In vitro Pex5 export assay, co-immunoprecipitation, RNAi knockdown, PEX5 stability assay, domain mapping |
Traffic (Copenhagen, Denmark) |
High |
21980954
|
| 2015 |
Cryo-EM structures of the Pex1/Pex6 complex in different nucleotide states reveal an unprecedented heterohexameric double ring in which Pex1 and Pex6 alternate; the N-terminal D1 ring is catalytically inactive and symmetric, while the C-terminal D2 ring is active and asymmetric. N1 of Pex1 is mobile; Pex6 N1 is packed against the ring. The architecture resembles p97/NSF, suggesting an analogous role in peroxisomal protein import to p97 in ERAD. |
Cryo-electron microscopy, computational domain fitting (Monte Carlo placement + energy minimization), nucleotide-state comparisons |
Proceedings of the National Academy of Sciences of the United States of America |
High |
26170309
|
| 2015 |
PEX6 localizes to cilia of retinal photoreceptor cells and to apical extensions of secretory ameloblasts/odontoblasts in mice, as shown by immunofluorescence, linking peroxisome biogenesis to ciliary function in the retina. |
Immunofluorescence microscopy in mouse retinal and dental tissues |
Human mutation |
Medium |
26593283
|
| 2018 |
The yeast Pex1/Pex6 heterohexamer is a protein translocase that unfolds and processively threads substrates through its central pore in a pore-loop-dependent and ATP-hydrolysis-dependent manner. Pex15 (the peroxisomal membrane tether) is recruited via its C-terminal disordered region engaging Pex1/Pex6 pore loops after first binding the N-terminal domains of Pex6. Pex15 also directly binds the cargo receptor Pex5, linking Pex1/Pex6 to the import machinery. |
In vitro unfolding/threading assay, cryo-EM structural analysis of Pex15–Pex1/Pex6 complex, pore-loop mutagenesis, ATP hydrolysis assay, co-immunoprecipitation (Pex15–Pex5 binding) |
Nature communications |
High |
29321502
|
| 2018 |
DTM-embedded monoubiquitinated PEX5 (Ub-PEX5) directly interacts with both PEX1 and PEX6 through its ubiquitin moiety, and the PEX5 polypeptide is globally unfolded during ATP-dependent extraction by the PEX1–PEX6 complex, establishing Ub-PEX5 as a bona fide substrate of the PEX1–PEX6 translocase. |
Cell-free in vitro extraction system, photoaffinity cross-linking, protein PEGylation assay |
The Journal of biological chemistry |
High |
29884772
|
| 2014 |
Deficiency of the exportomer components Pex1, Pex6, and Pex15 in S. cerevisiae causes enhanced pexophagy dependent on Atg11 and the pexophagy receptor Atg36; accumulation of ubiquitinated receptors at the peroxisomal membrane in pex1Δ/pex6Δ cells does not drive pexophagy in yeast (negative result for that trigger). Nearly all peroxisomal membranes associate with phagophore assembly sites in pex1Δ atg1Δ cells. |
Genetic screen of peroxisome import mutants, fluorescence microscopy, epistasis analysis (atg1Δ, atg11Δ, atg36Δ double mutants) |
Autophagy |
Medium |
24657987
|
| 2002 |
In PEX6-deficient CHO cells, peroxisomal ghosts are complex membrane structures (a central spherical body with double-membraned loops, with ER alongside); upon complementation with PEX6 cDNA, these structures become import-competent and are converted into functional peroxisomes, with catalase and acyl-CoA oxidase first accumulating in the lumen of the double-membraned loops. |
Genetic complementation with PEX6 cDNA, electron microscopy, immunofluorescence, biochemical fractionation |
Molecular biology of the cell |
Medium |
11854424
|
| 2007 |
In yeast, PEX6 (multicopy) suppresses aging defects in an atp2 mutant by improving mitochondrial import kinetics of Atp2p, requiring ATP binding/hydrolysis activity of Pex6p and specific residues in Atp2p; this reveals a novel role for Pex6p in mitochondrial maintenance and age asymmetry. |
Multicopy suppressor screen, epistasis analysis, mitochondrial import assay, ATP binding/hydrolysis mutants |
Aging cell |
Medium |
17465979
|
| 2023 |
Cryo-EM structures of yeast Pex1/Pex6 with an endogenous substrate trapped in the D2 ring central pore reveal that pairs of Pex1/Pex6 D2 subdomains engage substrate via a staircase of pore-1 loops with distinct properties; the inactive D1 ring undergoes conformational changes (widening/narrowing) fueled by D2 ATP hydrolysis; a 'twin-seam' Pex1/Pex6 D2 heterodimer disengages from the staircase to propagate mechanical force in a manner unique to heteromeric AAA-ATPases. |
Cryo-EM with endogenous substrate trap, structural analysis of nucleotide states |
Nature communications |
High |
37741838
|
| 2023 |
The N1 domain of Pex6 is required for Pex1/Pex6 function in vivo at the peroxisome, even though ΔN1-Pex6 retains ATPase activity in vitro. Crystal structure of the Pex6 N1 domain reveals the same fold as N-terminal domains of PEX1, CDC48, and NSF. Biochemical assays show Pex6 N1 mediates binding to the peroxisomal membrane tether Pex15 and also contacts an extended loop of the Pex1 D2 ATPase domain that stabilizes the heterohexamer. |
X-ray crystallography of isolated Pex6 N1 domain, cryo-EM reconstruction of Pex1/Pex6, AlphaFold2 modeling, in vitro ATPase assay, biochemical binding assays, in vivo functional complementation |
The Journal of biological chemistry |
High |
38036174
|
| 2003 |
In Hansenula polymorpha, a point mutation G737E in the first AAA module of Pex6p confers cold-sensitive peroxisome biogenesis; a second intragenic mutation R1000G in the second AAA domain acts as an intragenic suppressor restoring growth at permissive temperature. These residues in both AAA domains are important for Pex6p function. |
UV mutagenesis, intragenic suppressor analysis, electron microscopy of peroxisome formation |
FEMS yeast research |
Medium |
14613878
|
| 2000 |
A temperature-sensitive missense mutation L57P in PEX6 (but not the counterpart L111P in PEX1, nor G708D in PEX6) causes temperature-sensitive peroxisome biogenesis in patient fibroblasts and CHO transfectants, demonstrating that L57 in the N-terminal region of PEX6 is critical for its activity and that temperature-sensitive residues are not conserved between PEX6 and PEX1. |
Temperature-shift experiments in patient fibroblasts and CHO transfectants, morphological and biochemical peroxisome assays |
Pediatric research |
Medium |
11004248
|
| 2026 |
Pex6 (together with Pex26) regulates reversible, ATP-dependent topological remodeling of the peroxisomal membrane protein Pex14: under normal conditions Pex14's N-terminal domain faces the lumen, but loss of Pex6 or Pex26, or pharmacological inhibition of AAA+ ATPases, causes the Pex14 N-terminus to become cytoplasm-facing. Inhibition of ubiquitin activation (blocking Pex5 ubiquitination and extraction) prevented this reorientation, placing Pex5 ubiquitination and Pex6-mediated extraction upstream of Pex14 topological remodeling. |
Immunofluorescence microscopy, protease protection assay, pharmacological inhibitors (AAA+ ATPase inhibitor, MLN-7243 ubiquitin E1 inhibitor), Pex6/Pex26 deficiency cell lines |
The Journal of biological chemistry |
Medium |
41581879
|